Human umbilical cord blood cells transfected with VEGF and L(1)CAM do not differentiate into neurons but transform into vascular endothelial cells and secrete neuro-trophic factors to support neuro-genesis-a novel approach in stem cell therapy.
Genetically modified mono-nuclear cell fraction from human umbilical cord blood (HUCB) expressing human vascular endothelial growth factor (VEGF) and mouse neural L(1) cell adhesion molecule (L(1)CAM) were used for gene-stem cell therapy of transgenic (G)93(A) mice adopted as an animal amyotrophic lateral sclerosis (ALS) model. We generated non-viral plasmid constructs, expressing human VEGF(165) (pcDNA-VEGF) and mouse neural L(1) cell adhesion molecule (pcDNA-mL(1)CAM). Mono-nuclear fraction of HUCB cells were transiently transfected by electro-poration with a mixture of expression plasmids (pcDNA-VEGF+pcDNA-mL(1)CAM). Sixteen transgenic female and male mice were randomly assigned to three groups: (1) transplantation of genetically modified HUCB cells expressing L(1) and VEGF (n=6), (2) transplantation of un-transfected HUCB cells (n=5), and (3) control group (n=5). In first two experimental groups 1x10(6) cells were injected retro-orbitally in pre-symptomatic 22-25-week-old (G)93(A) mice. Our results demonstrate that HUCB cells successfully grafted into nervous tissue of ALS mice and survived for over 3 months. Therefore, genetically modified HUCB cells migrate in the spinal cord parenchyma, proliferate, but instead of transforming into nerve cells, they differentiate into endothelial cells forming new blood vessels. We propose that: (A) expression of mouse neural L(1)CAM is responsible for increased homing and subsequent proliferation of transplanted cells at the site of neuro-degeneration, (B) expression of human VEGF directs HUCB cell differentiation into endothelial cells, and (C) neuro-protective effect may stem from the delivery of various neuro-trophic factors from newly formed blood vessels.
Rizvanov
Albert A
AA
Department of Histology, Kazan State Medical University, ul. Butlerova 49, R-420012 Kazan, Russia.
Kiyasov
Andrey P
AP
Gaziziov
Ilnaz M
IM
Yilmaz
Tatyana S
TS
Kaligin
Maxim S
MS
Andreeva
Dina I
DI
Shafigullina
Aigul K
AK
Guseva
Darya S
DS
Kiselev
Sergey L
SL
Matin
Keyvan
K
Palotás
András
A
Islamov
Rustem R
RR
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism*; D005260:Female; D005822:Genetic Vectors; D006085:Graft Survival / Q000502:physiology; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D018919:Neovascularization, Physiologic / Q000235:genetics*; D009414:Nerve Growth Factors / Q000378:metabolism*; D039842:Neural Cell Adhesion Molecule L1 / Q000235:genetics; D055495:Neurogenesis / Q000502:physiology*; D013116:Spinal Cord / Q000166:cytology / Q000502:physiology; D033581:Stem Cell Transplantation / Q000379:methods*; D014162:Transfection / Q000379:methods; D016896:Treatment Outcome; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics
D009414:Nerve Growth Factors; D039842:Neural Cell Adhesion Molecule L1; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A